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Evaluation of ABCG 2 and p63 expression in canine cornea and cultivated corneal epithelial cells
Author(s) -
Morita Maresuke,
Fujita Naoki,
Takahashi Ayaka,
Nam Eun Ryel,
Yui Sho,
Chung Cheng Shu,
Kawahara Naoya,
Lin Hsing Yi,
Tsuzuki Keiko,
Nakagawa Takayuki,
Nishimura Ryohei
Publication year - 2015
Publication title -
veterinary ophthalmology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.594
H-Index - 50
eISSN - 1463-5224
pISSN - 1463-5216
DOI - 10.1111/vop.12147
Subject(s) - cornea , corneal epithelium , pathology , biology , immunohistochemistry , epithelium , immunocytochemistry , corneal diseases , medicine , neuroscience
Objective To examine the expressions of ABCG 2 and p63 in canine corneal epithelia and to evaluate their significance in corneal regeneration. Procedures Canine corneal and limbal epithelial cells were obtained from five healthy beagle dogs. We analyzed the morphological properties of cultivated limbal and corneal epithelial cells. We compared the expressions of ABCG 2 and p63 in the limbus and central cornea by immunohistochemistry and real‐time quantitative PCR . We analyzed the expression of these markers in cultivated cells by immunocytochemistry and real‐time quantitative PCR . Results The limbal epithelial cells were smaller and proliferated more rapidly than the corneal epithelial cells in primary cultures. The corneal cells failed to be subcultured, whereas the limbal cells could be subcultured with increasing cell size. ABCG 2 was localized in the basal layer of the limbal epithelium, and p63 was widely detected in the entire corneal epithelia. ABCG 2 expression was significantly higher, and p63 was slightly higher in the limbus compared with the central cornea. ABCG 2 was detected only in limbal cells in primary culture, not in corneal cells or passaged limbal cells. p63 was detected in both limbal and corneal cells and decreased gradually in the limbal cells with the cell passages. Conclusions ABCG 2 was localized in canine limbal epithelial cells, and p63 was widely expressed in canine corneal epithelia. ABCG 2 and p63 could prove to be useful markers in dogs for putative corneal epithelial stem cells and for corneal epithelial cell proliferation, respectively.